KR20220155132A - Composition for selective delivery of drugs comprising encapsulated, cordycepin-inserted hydrotalcite - Google Patents

Abstract

The present invention relates to a method for manufacturing encapsulated and cordycepin-inserted hydrotalcite which comprises the following steps: removing H_2O and CO_3 from hydrotalcite; inserting cordycepin into the hydrotalcite from which H_2O and CO_3 are removed; and encapsulating by treating the surface of the hydrotalcite into which cordycepin is inserted with polysaccharide.

Description

Composition for selective delivery of drugs containing encapsulated cordycepin-inserted hydrotalcite

The present invention relates to a composition for the selective delivery of a drug comprising an encapsulated cordycepin-intercalated hydrotalcite.

Cordyceps sinensis, which exhibits anti-immune properties, is a medicinal mushroom that grows on insects as hosts. It originates from the fact that it parasitizes on insects in winter and grows like grass in summer to make mushrooms. have.

As for the representative efficacy of cordyceps, many efficacies such as anticancer action, immune action, improvement of liver function, improvement of high blood pressure, aging, diabetes, heart disease, and nutritional tonic have been reported. It contains various ingredients such as systemic substances, showing efficacy. Therefore, a number of foods, cosmetics, or medicines using Cordyceps sinensis have been studied. For example, Korean Patent Publication No. 10-2021-0020633 discloses a cosmetic composition containing an extract of Cordyceps sinensis, and Korean Patent Publication No. 10-2021-0001396 discloses a method for preparing a patch using Cordyceps sinensis powder.

On the other hand, hydrotalcite is a substance used as an antacid in the Korean Pharmacopoeia and has a double layered structure.

It is an object of the present invention to prepare materials that can be used in compositions for drug delivery.

Another object of the present invention is to provide a composition for drug delivery.

In order to achieve the above object, the present invention,

removing H ₂ O and CO ₃ from hydrotalcite;

inserting cordycepin into the hydrotalcite from which the H ₂ O and CO ₃ are removed; and

Encapsulation by treating the surface of the hydrotalcite into which cordycepin is inserted with polysaccharide,

A method for preparing encapsulated cordycepin-incorporated hydrotalcite is provided.

Also, the present invention

A drug delivery composition comprising the encapsulated cordycepin-intercalated hydrotalcite of the present invention is provided.

Also, the present invention

A pharmaceutical composition comprising the composition for drug delivery is provided.

The encapsulated cordycepin-inserted hydrotalcite of the present invention prevents drug degradation by enzymes and other substances during drug delivery and releases cordycepin and piperazine-based substances through acid dissolution and anion exchange in the intestine. By protecting the cordycepin substance from deamination by adenosine deaminase (ADA), the anti-immune performance can be maximized.

FIG. 1 confirms XRD analysis results of samples surface-treated with polysaccharide in Example 1, after hydrotalcite synthesis, before thermal ionization, after heat treatment, and after inserting cordycepin into hydrotalcite. 1 describes step 1 of Example 1 and shows the change in the distance between layers of hydrotalcite for each process. The upper figure of FIG. 1 shows the contents of XRD taken after three processes for hydrotalcite, and the lower figure shows the change in the distance of the two-layer structure after three processes by enlarging only the XRD 003 part showing the distance between the two-layer phases of hydrotalcite. . The arrow indicates that part 003 has been enlarged.
2 is a result of confirming the content of cordycepin in material (c) isolated from Cordyceps sinensis in step 2 of Example 1 using HPLC.
3 is a result of confirming the content of cordycepin in material (c) isolated from Cordyceps sinensis in step 2 of Example 1 using H-NMR
4 is a result of confirming the content of cordycepin in the material (c) isolated from Cordyceps sinensis in Step 2 of Example 1 using C-NMR.
5 is a result of confirming cell death for cytotoxicity using liver organoids. In Table 4, the conditions for elution from 0.1 N-HCl in the small intestine and absorption into the body are the ratios of cordycepin (1) and polysaccharide (3). After mixing the cordycepin of Examples 1-3 and the polysaccharide 1-4 at such a mixing ratio, a liver organoid test was conducted at concentrations of 3.14 ppm, 6.28 ppm, and 12.56 ppm.
6 shows the results of observing the size and viability of liver organoids using the samples of Examples 1 to 3. At this time, the y-axis represents the size of liver organoid cells, and 1 is the standard size ratio (%). Figure 6 shows the size of liver organoid cells, and the size ratio (%), and the size of the cells as well as the survival rate can be confirmed by the large and small bar sizes in the graph. That is, when the bar graph becomes smaller, cells are dying.

The present invention,

removing H ₂ O and CO ₃ from hydrotalcite;

inserting cordycepin into the hydrotalcite from which the H ₂ O and CO ₃ are removed; and

Encapsulation by treating the surface of the hydrotalcite into which cordycepin is inserted with polysaccharide,

It relates to a method for preparing encapsulated cordycepin-incorporated hydrotalcite.

Also, the present invention

The present invention relates to a composition for drug delivery comprising encapsulated cordycepin-intercalated hydrotalcite.

Also, the present invention

It relates to a pharmaceutical composition comprising the composition for drug delivery.

Also, the present invention

The present invention relates to a cordycepin delivery composition comprising encapsulated cordycepin-intercalated hydrotalcite.

Also, the present invention

It relates to a pharmaceutical composition comprising the composition for delivering cordycepin.

Also, the present invention

It relates to a pharmaceutical composition comprising the composition for delivering cordycepin.

Also, the present invention

It relates to a food composition comprising the cordycepin delivery composition.

In addition, the present invention relates to a method for extracting high-content cordycepin from Cordyceps sinensis.

In addition, the present invention is inserted between the middle of the middle of the material of hydrotalcite so that it can be delivered to the intestine without being easily decomposed in the stomach when drinking, and the surface is treated with polysaccharide extracted from Cordyceps sinensis, and after safely transported, the cordycepin component is delivered and absorbed by ion exchange in the intestine. The present invention relates to a composition that can be used as a seasoning food or health functional food as well as a physiologically active natural medicine as a substance having an anti-immune enhancing function in the body.

Hereinafter, the present invention will be described in detail.

H in hydrotalcite ₂ O and CO ₃ steps to remove

The method of preparing the encapsulated cordycepin-incorporated hydrotalcite of the present invention includes removing H ₂ O and CO ₃ from the hydrotalcite.

The step of removing H ₂ O and CO ₃ from hydrotalcite is

(i) adding filtrate B and filtrate A to a basic aqueous solution, reacting the mixture, and dehydrating the mixture to obtain a solid content. At this time, the method for preparing the filtrate A may include adding aluminum hydroxide (Al(OH) ₃ ), NaOH, and water and heating. Al(OH) ₃ and NaOH may be used in a weight ratio of 1:1.5 to 5.

In this case, the method for preparing the filtrate B may include dissolving Mg(Cl) ₂ 6H ₂ O in water. Preferably, it can be prepared by dissolving Mg(Cl) ₂ 6H ₂ O in water and then filtering.

Hydrotalcite is a micro-sized hybrid material, which is used in the present invention to prepare a composition for drug delivery, preferably drug selective delivery. Hydrotalcite has a double layered structure, and the present invention uses hydrotalcite having such a double layered structure to make a layered material with a size of 150 to 250 microns, and the cordyceps extract cordycepin substance inside the layered structured material. is inserted, and the surface is treated with polysaccharides, which are abundant in Cordyceps sinensis and other mushrooms, to prevent decomposition from enzymes and other substances during transportation to Biomolecules, and cordycepin and piperazine-based substances are produced through acid dissolution and anion exchange in the intestine. It is released to protect the cordycepin substance from deamination by adenosine deaminase (ADA) to make the most of its anti-immune performance.

Cordycepin Remind H ₂ O and CO ₃ Step of inserting into hydrotalcite from which is removed

The method of preparing encapsulated cordycepin-incorporated hydrotalcite of the present invention includes inserting cordycepin into the hydrotalcite from which H ₂ O and CO ₃ are removed.

The cordycepin may be Cordyceps-derived cordycepin. Cordyceps sinensis of the present invention may be Sinensis Cordyceps sinensis, Militaris Cordyceps sinensis, Silkworm Cordyceps sinensis, Cordyceps sinensis of various insects, and preferably Militaris Cordyceps sinensis. Preferably, as the cordycepin material, a material obtained by selectively extracting a material contained in Cordyceps sinensis and isolating cordycepin may be used.

In the step of inserting cordycepin into the hydrotalcite from which H ₂ O and CO ₃ are removed, the cordycepin is in the form of a cordycepin-containing material, wherein the material contains 90% by weight or more of cordycepin. can

Hydrotalcite and cordycepin may be used in a weight ratio of 100:0.0001 to 0.05, and cordycepin can be effectively incorporated into hydrotalcite within the above range.

In the present invention, the layer size of the single structure of micro-sized hydrotalcite is provided to be 7.0 Å to 9.0 Å, heat treatment is performed at 350 ° C to 420 ° C to change the size of the layer structure to 6.5 Å to 7.8 Å, and then When the cordycepin substance is inserted, the size of the layered structure is transformed from 7.0Å to 18.0Å and inserted into the double layered structure.

Encapsulation by treating the surface of the hydrotalcite into which cordycepin is inserted with polysaccharide

The method of preparing encapsulated cordycepin-incorporated hydrotalcite of the present invention includes encapsulating the surface of the cordycepin-incorporated hydrotalcite by treating it with a polysaccharide. The polysaccharide may be a polysaccharide derived from Cordyceps sinensis. Preferably, a polysaccharide obtained by selectively extracting a substance contained in Cordyceps sinensis and isolating the polysaccharide may be used. Additionally, as the polysaccharide, a material containing β-glucan and dextrin, which are polysaccharides obtained from Cordyceps sinensis, may be used, and these materials may be selectively mixed and used.

At this time, the hydrotalcite and the polysaccharide may be used in a weight ratio of 100:100 to 500, and within this range, the hydrotalcite into which cordycepin is inserted can be effectively coated with the polysaccharide and encapsulated.

Method for preparing encapsulated, cordycepin-incorporated hydrotalcite

The average particle size of the encapsulated cordycepin-intercalated hydrotalcite of the present invention may be 0.5 to 15 μm. The encapsulated cordycepin-inserted hydrotalcite of the present invention inserts a drug, preferably cordycepin, into the layered structure of the hydrotalcite to prevent decomposition due to enzymes and other substances during transport and acid dissolution in the intestine. In addition, cordycepin and piperazine-based substances are released through anion exchange to protect cordycepin substances from deamination by adenosine deaminase (ADA), so that the anti-immune performance can be maximized. In addition, it can be sufficiently utilized as a delivery medium for drinking and food additives and further biopharmaceuticals by utilizing it, and it can be used as a selective drug delivery material for hybrid materials by utilizing hydrotalcite containing cordycepin having an anti-immunity effect.

The substance obtained from the cordyceps sinensis extract is a substance derived from natural products and is widely used for medicinal purposes. There is very little concern about side effects or safety, so when used as food or health functional food through oral intake or even as a medicine derived from natural substances, the development of this technology is It is possible to exert the efficacy and effectiveness of prevention or treatment, and it is judged that enough time can be secured for the cordycepin substance to act in the body.

The encapsulated cordycepin-inserted hydrotalcite of the present invention is characterized in that the cordycepin material is inserted into the layered structure of the micro-sized hydrotalcite and then the surface is treated with cordyceps polysaccharide. It is possible to obtain a cordyceps extract liquid composition for the purpose of incorporating the cordycepin substance into the living body by ion exchange. Therefore, by using the encapsulated cordycepin-inserted hydrotalcite of the present invention, a selective composition necessary for immune function improvement, prevention, treatment, etc. is provided.

That is, the present invention is used as an antacid in the Korean Pharmacopoeia and by using the characteristics of the drug, the area of the double layered structure is widened to the interlayer area of the existing hydrotalcite, so that cordycepin substances can be more easily inserted, and the polysaccharide contained in Cordyceps sinensis is more easily Surface treatment with phosphorus and dextrin-containing substances to make a micro-sized liquid substance, which makes it easier to eat when drinking. The immune functional composition can be used as drinking water, other foods, health functional foods, and even medicines. It is to make a composition that can be easily used so that anyone can use it easily.

In one aspect of the present invention, the present invention is a material containing β-glucan, a polysaccharide obtained from Cordyceps sinensis extraction, and dextrin to easily obtain a powder in a material in which cordycepin material is inserted into the layered structure of micro-sized hydrotalcite. Surface treatment to provide a micro-sized liquid composition or powder composition encapsulated.

Composition for drug delivery

The present invention relates to a drug delivery composition comprising the encapsulated cordycepin-intercalated hydrotalcite of the present invention.

In this case, the drug may be cordycepin. In one aspect of the present invention, the present invention may be directed to a composition for delivery of cordycepin comprising an encapsulated, cordycepin-intercalated hydrotalcite of the present invention.

medicinal composition

The present invention relates to a pharmaceutical composition comprising the composition for drug delivery.

The present invention relates to a pharmaceutical composition comprising the composition for delivery of cordycepin.

The pharmaceutical composition may include 0.01 to 80% by weight, preferably 0.02 to 65% by weight of the drug delivery composition, the cordycepin delivery composition, or the encapsulated cordycepin-inserted hydrotalcite. can do. However, this may be increased or decreased according to the needs of the administering person, and may be appropriately increased or decreased depending on the circumstances such as the age, dietary habits, nutritional status, and progress of the disease of the subject.

The pharmaceutical composition of the present invention can be administered orally or parenterally and can be used in the form of general pharmaceutical preparations. Preferred pharmaceutical preparations include preparations for oral administration such as tablets, hard or soft capsules, solutions, suspensions, and the like, and these pharmaceutical preparations are pharmaceutically acceptable carriers, for example, excipients in the case of preparations for oral administration, It can be prepared using a binder, disintegrant, lubricant, solubilizer, suspending agent, preservative or bulking agent.

The dosage of the drug delivery composition of the present invention, the cordycepin delivery composition, or the encapsulated pharmaceutical composition containing the cordycepin-inserted hydrotalcite may vary depending on the patient's condition, age, gender, complication, etc. Although it can be determined by experts according to various factors, it is generally administered at a dose of 0.1 mg to 10 g, preferably 10 mg to 5 g per 1 kg of adult. In addition, the daily dose of the pharmaceutical composition per unit dosage form or a dose of 1/2, 1/3 or 1/4 thereof may be contained, and may be administered 1 to 6 times a day. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be less than the above range, and the active ingredient may be used in an amount greater than the above range because there is no problem in terms of safety.

food composition

The present invention relates to a food composition comprising the composition for delivering cordycepin. Also, the present invention relates to a food composition comprising the encapsulated cordycepin-intercalated hydrotalcite of the present invention. The food includes food additives.

The food of the present invention may be a health supplement, health functional food, functional food, etc., but is not limited thereto, and includes natural food, processed food, and general food materials containing the cordyceps sinensis extract of the present invention.

In the food composition of the present invention, the drug delivery composition, the cordycepin delivery composition, or the encapsulated cordycepin-inserted hydrotalcite may be added as is or used together with other foods or food compositions. It can be used appropriately depending on the method. The mixing amount of the active ingredient can be suitably determined depending on the purpose of its use (prevention, improvement or therapeutic treatment). In general, the cordyceps sinensis extract of the present invention may be added in an amount of 0.1 to 5% by weight, preferably 2 to 3% by weight, based on 100% by weight of the raw material of the food or beverage when preparing food or beverage. The effective dose of the drug delivery composition of the present invention, the cordycepin delivery composition, or the encapsulated cordycepin-inserted hydrotalcite may be used according to the effective dose of the pharmaceutical composition, but health and In the case of long-term intake for the purpose of hygiene or health control, it may be less than the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety.

There is no particular limitation on the type of food. The food composition may be used in the form of formulations for oral administration such as tablets, hard or soft capsules, solutions, suspensions, etc. It can be prepared using a binder, disintegrant, lubricant, solubilizer, suspending agent, preservative or bulking agent.

Examples of the drug delivery composition of the present invention, the cordycepin delivery composition, or the encapsulated food to which the cordycepin-inserted hydrotalcite can be added include meat, sausage, bread, chocolate, candy, and snacks. , confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages and vitamin complexes, and other nutritional supplements, but not limited to these types of foods. .

feed composition

The present invention relates to a feed composition comprising the cordycepin delivery composition. Also, the present invention relates to a feed composition comprising the encapsulated cordycepin-incorporated hydrotalcite of the present invention.

The feed composition of the present invention contains 0.01 to 5.00% by weight of the drug delivery composition, the cordycepin delivery composition, or the encapsulated cordycepin-inserted hydrotalcite, preferably 0.03 to 3.00 wt%, based on dry matter. % by weight, more preferably 0.05 to 0.10% by weight.

The feed composition of the present invention may be a feed composition for vertebrates such as cows and pigs, poultry such as chickens and ducks, fish, and crustaceans, but is not limited thereto. The feed composition of the present invention can be used regardless of the mixing ratio in the feed composition. For example, there is no problem in using the present invention even if the feed has a high protein content.

Drug delivery method

The present invention relates to a method for delivering a drug in the body of a subject, comprising the step of administering the drug delivery composition, the cordycepin delivery composition, or the encapsulated composition containing cordycepin-inserted hydrotalcite to the subject. it's about how

The subject may be diagnosed or subjectively or objectively determined to have a benefit or need to receive a drug, or may be a subject. The subject may be a mammal including a human.

Advantages and features of the present invention, and how to achieve them, will become clear with reference to the detailed description of the following embodiments. However, the present invention is not limited to the embodiments disclosed below and will be implemented in various forms different from each other, only these embodiments make the disclosure of the present invention complete, and common knowledge in the art to which the present invention pertains. It is provided to completely inform the person who has the scope of the invention, and the present invention is only defined by the scope of the claims.

<Materials and methods>

Cordyceps militaris, which was grown using brown rice, was used for Cordyceps militaris. Reagent dextrin, a food additive, was purchased and used as a commercially available product. Hereinafter, in the experimental examples, samples were prepared and experiments were performed under conditions in which cordycepin was absorbed in the small intestine via the stomach.

<Example 1>

[Step 1]

Micro-sized hydrotalcite was synthesized through the following process, and H ₂ O and carbonate (CO ₃ ) were removed from the inside of the layered structure. First, put 15.63 g of 99.8% Al(OH) ₃ as a reagent, 48.98 of 48.98% NaOH as a reagent, and 50mL of distilled water into a 250 ml beaker and raise the temperature to near the boiling point until Al(OH) ₃ is completely dissolved. After heating, the insoluble matter was filtered out with filter paper to prepare filtrate A. On the other hand, 300mL of distilled water was put in a 500mL beaker, 124.47g of 98% Mg(Cl) ₂ 6H ₂ O was completely dissolved, and filtrate B was prepared by filtering out the insoluble matter with filter paper.

Then, in a 3L beaker, completely dissolve 49.98g of reagent 98% NaHCO ₃ in 2L of water, filter out the insoluble content with filter paper, add the prepared filtrate B to the received filtrate, and slowly add the prepared filtrate A while stirring with a homogenizer at 7500 rpm. After hydrothermal synthesis reaction at 170 ° C for 4 hours in a High Pressure Reactor, dehydration with filter paper, the solid content was put into 3L of distilled water, milled for 4 hours in a stirrer with 0.2mm beads, and then dehydrated to synthesize hydrotalcite. Thereafter, by drying at 350 ° C to 420 ° C to remove H ₂ O and CO ₃ , 34.4 g of average particle size 0.1 to 0.3 micro hydrotalcite was obtained, and the analysis result was confirmed by XRD measurement in FIG.

[Step 2]

Cordycepin material was isolated from Cordyceps sinensis as follows. 1.5 kg of dried Cordyceps sinensis powder was added to 15 L of 50 v/v% ethanol, extracted twice, and the obtained filtrate was concentrated by darkening to obtain 183.7 g of a substance (a) containing 50 v/v% ethanol and 11.1% cordycepin. , Using HP-20 column chromatography, at this time, using 20 v / v% ethanol as a mobile phase to purify and fractionate the material (a) to obtain 39.44 g of material (b) containing 51.7 wt% cordycepin, In order to increase the content of cordycepin, the material (b) was separated into a filtrate and a supernatant using 20 v/v% ethanol as a mobile phase and centrifuged. The filtrate was cooled and dried to separate cordycepin therefrom to obtain 18.36 g of material (c) containing cordycepin with a final purity of 97%, which was analyzed to confirm cordycepin in the material (c) by HPLC in FIG. 2. And the content of cordycepin was confirmed by H-NMR of FIG. 3 and C-NMR of FIG.

[Step 3]

Polysaccharides isolated from Cordyceps sinensis were prepared as follows. The supernatant obtained in step 2 was used. 100 mL of distilled water was added to the precipitate using the supernatant for fractionation of cordycepin, put into a dialysis bag, put in a refrigerator, and dialyzed at 3° C. for 24 hours. At this time, the solvent is separated from the supernatant using a dialysis membrane in the dialysis bag and crystallization proceeds. After dialysis, a crystal substance mixed with some solvent remains in the dialysis bag. , and dried at 50° C. for 24 hours to obtain 108.0 g of polysaccharide.

[Step 4]

Cordycepin isolated from Cordyceps sinensis was inserted into the layered structure of micro-sized hydrotalcite. First, 100 mL of distilled water was placed in a 250 mL beaker, and 10 g of hydrotalcite prepared in step 1 was added thereto. While stirring this with a homogenizer at 7500 rpm, 0.5024 mg of cordycepin prepared in step 2 was added and stirred for 30 minutes to insert cordycepin into the hydrotalcite bilayer structure. At this time, the amount of cordycepin inserted into the bilayer structure of hydrotalcite was 50.24 ppm.

[Step 5]

The cordycepin-incorporated hydrotalcite was encapsulated by final surface treatment with polysaccharide. 24.0 g of the polysaccharide isolated in step 3 and 6.0 g of food additive dextrin (OCI formula (C ₆ H ₁₀ O ₅ ) n.nH ₂ O) were added to the cordycepin-inserted hydrotalcite obtained in step 4, and then 60 A sample containing encapsulated cordycepin-incorporated hydrotalcite was prepared by stirring for a minute.

[Step 6]

Stirring was stopped in step 5 to obtain encapsulated cordycepin hydrotalcite, which was filtered and lyophilized to prepare the encapsulated cordycepin hydrotalcite sample of Example 1.

The encapsulated cordycepin hydrotalcite sample was analyzed through XRD measurement in FIG. 1 .

<Example 2>

Steps 1 to 3 were performed in the same manner as in Example 1.

[Step 4]

Cordycepin isolated from Cordyceps sinensis was inserted into the layered structure of micro-sized hydrotalcite. First, 100 mL of distilled water was put into a 250 mL beaker, and 10 g of hydrotalcite prepared in step 1 was added thereto. While stirring this at 7500 rpm, the homogenizer, 0.2512 mg of cordycepin prepared in step 2 was added and stirred for 30 minutes to insert cordycepin into the hydrotalcite bilayer structure. At this time, the amount of cordycepin inserted into the bilayer structure of hydrotalcite was 25.12 ppm.

[Step 5]

The cordycepin-incorporated hydrotalcite was encapsulated by final surface treatment with polysaccharide. 24.0 g of the polysaccharide isolated in step 3 and 6.0 g of food additive dextrin (OCI formula (C ₆ H ₁₀ O ₅ ) n.nH ₂ O) were added to the cordycepin-inserted hydrotalcite obtained in step 4, and then 60 A sample containing encapsulated cordycepin-embedded hydrotalcite was prepared by stirring for 10 minutes.

[Step 6]

Stirring was stopped in step 5, and encapsulated cordycepin hydrotalcite was obtained, filtered, and lyophilized to prepare an encapsulated cordycepin hydrotalcite sample of Example 2.

<Example 3>

Steps 1 to 3 were performed in the same manner as in Example 1.

[Step 4]

Cordycepin isolated from Cordyceps sinensis was inserted into the layered structure of micro-sized hydrotalcite. First, 100 mL of distilled water was put into a 250 mL beaker, and 10 g of hydrotalcite prepared in step 1 was added thereto. While stirring this at 7500 rpm, the homogenizer, 0.1256 mg of cordycepin prepared in step 2 was added and stirred for 30 minutes to insert cordycepin into the hydrotalcite bilayer structure. At this time, the amount of cordycepin inserted into the bilayer structure of hydrotalcite was 12.56 ppm.

[Step 5]

The cordycepin-incorporated hydrotalcite was encapsulated by final surface treatment with polysaccharide. 24.0 g of the polysaccharide isolated in step 3 and 6.0 g of food additive dextrin (OCI formula (C ₆ H ₁₀ O ₅ ) n.nH ₂ O) were added to the cordycepin-inserted hydrotalcite obtained in step 4, and then 60 A sample containing encapsulated cordycepin-incorporated hydrotalcite was prepared by stirring for a minute.

[Step 6]

Stirring was stopped in step 5 to obtain encapsulated cordycepin hydrotalcite, which was filtered and lyophilized to prepare the encapsulated cordycepin hydrotalcite sample of Example 3.

<Comparative Example 1>

12.5 ppm of cordycepin was added to 10.0 g of cellulose, stirred with a homogenizer at 7500 rpm, and nanospray coacervation was performed to encapsulate cordycepin containing cordycepin to prepare a sample in which cordycepin was encapsulated with cellulose.

The samples prepared in Examples 1 to 3 and Comparative Example 1 are summarized in Table 1 below.

Cordycepin (mg) Carrier (g) Surface treatment agent (g) Example 1 0.5024 Hydrotalcite (10g) Cordyceps sinensis extract-derived polysaccharide (24.0g) and dextrin (6.0g) Example 2 0.2512 Hydrotalcite (10g) Cordyceps sinensis extract-derived polysaccharide (24.0g) and dextrin (6.0g) Example 3 0.13 Hydrotalcite (10g) Cordyceps sinensis extract-derived polysaccharide (24.0g) and dextrin (6.0g) Comparative Example 1 12.5 ppm by weight of cellulose - Cellulose (10.0 g)

<Experimental Example 1> XRD measurement results

For the samples of Examples 1 to 3 and Comparative Example 1, XRD was measured using an X-ray diffractometer (XRD) to confirm the form in which cordycepin was inserted into the hydrotalcite bilayer structure. At this time, 2Theta of the measurement range was measured from 5° to 70°, and the size change was measured before heat treatment of hydrotalcite, after heat treatment, and after insertion of cordycepin into the layered structure (distance 003 between the two-layered hydrotalcite structure). Measured.

It was confirmed that the size of the bilayer structure of hydrotalcite changes according to the insertion concentration of cordycepin, and through this, it was confirmed that not only cordycepin but also other drugs can be inserted into hydrotalcite and used. In Table 2, room temperature dehumidification means that after synthesizing hydrotalcite in a wet manner, moisture is generally removed at a high temperature, and moisture is dried at room temperature (Table 2).

Before heat treatment (003) peak
(Å) After heat treatment (003) peak
(Å) (003) peak (Å) after surface treatment agent Example 1 7.56 6.80 23.33 Example 2 7.56 6.80 18.46 Example 3 7.56 6.80 12.10 Room temperature dehumidification in Preparation Example 1 7.56 6.80 7.78

<Experimental Example 2> Measurement of particle size and degree of dispersion

The particle size and dispersion of cordycepin hydrotalcite encapsulated in the samples of Examples 1 to 3 and Comparative Example 1 were measured. At this time, the particle size was measured using a Malvern mastersizer 2000 (Malvern instruments Ltd, Malvern UK, Accessory hydro 2000s), and the polydispersity index (PDI) was measured using a Malvern zeta nano ZS (Malvern instruments Ltd, Malvern UK). , At this time, ethanol was used as the dispersion solution. Dispersity (Polydispersity index (PDI)) is generally 0 < PDI < 0.2, and it can be said that the dispersion of particles is very good, and there is no unit.

As a result, the particle size and dispersion of cordycepin hydrotalcite encapsulated in the samples of Examples 1 to 3 were smaller than those of Comparative Example 1. Therefore, it was found that the particles of the encapsulated cordycepin hydrotalcite were smaller than those of Comparative Example 1, so the absorption rate was good in the intestine, and the amount of dissolution increased, and the dispersion stability was also more stable than that of Comparative Example 1 (Table 3).

Particle size (㎛) dispersion hydrotalcite particle size 0.20 to 0.35 submerged Example 1 2.30±0.2 0.18±0.01 Example 2 2.28±0.2 0.15±0.01 Example 3 2.26±0.2 0.14±0.01 Comparative Example 1 20.21±21 0.21±0.01

<Experimental Example 3> Evaluation of cordycepin release characteristics in gastrointestinal conditions and small intestine conditions

The cordycepin release characteristics of the samples of Examples 1 to 3 and Comparative Example 1 were evaluated under gastrointestinal conditions and small intestine conditions. Evaluation in the gastric condition was performed by using 1N-HCl in distilled water to create a condition of pH 1.5 of 10mL, and then leaving 5.0g of the sample there for 10 minutes, separating the filtrate and measuring the cordycepin content by HPLC. In the small intestine condition, the solid content (i.e., the hydrotalcite inserted with the encapsulated cordycepin prepared in Examples 1 to 3 used in the gastrointestinal condition and the cellulose capsule material in Comparative Example 1) was added to 10 mL of 0.1 N-HCl for 2 hours. to create small intestine conditions, separate the filtrate, and measure the content of cordycepin by HPLC.

As a result, the samples of Examples 1 to 3 showed a lower dissolution rate than Comparative Example 1 in the stomach condition, but a higher dissolution rate than Comparative Example 1 in the small intestine condition (Table 4).

Cordycepin content of 1N-HCl filtrate
(ppm) dissolution rate
(%) Cordycepin content of 0.1 N-HCl filtrate
(ppm) dissolution rate
(%) Example 1 0.39 3.12 11.75 94.00 Example 2 0.16 2.56 5.94 94.89 Example 3 0.06 1.91 3.02 95.04 Comparative Example 1 4.38 35.04 6.92 55.36

<Experimental Example 4> Evaluation of toxicity in liver organoids

<4-1>

Responsiveness and predictability of liver organoid toxicity through treatment with hepatotoxicity-inducing drugs based on the maximum dissolution rate were evaluated.

Troglitazone, Rac perhexiline maleate, and Thioridazine HCI, which are known to have existing liver toxicity, were added to liver organoids (differentiation 30 days, passage 2) produced by Organ Factory at various concentrations (Troglitazone: 20, 200, 400 uM, Rac After treatment with perhexiline maleate: 0.2, 2, 20 uM, Thioridazine HCI: 0.5, 5, 50 uM) for 24 hours, the size and shape of liver organoids were observed, and apoptosis was investigated through TUNEL assay. Responsiveness and predictability were evaluated by confirming apoptosis and cell viability for toxic drugs.

<4-2>

Drug toxicity of liver organoid-based samples was evaluated.

The condition for absorption into the body after being eluted from 0.1 N-HCl in the small intestine conditions of Table 4 is the ratio of cordycepin (1) to polysaccharide (3). Anticipating this, after mixing the cordycepin of Examples 1-3 and the polysaccharide 1-4, a liver organoid test was conducted at concentrations of 3.14 ppm, 6.28 ppm, and 12.56 ppm, respectively. That is, C1:P3 represents the mixing ratio of cordycepin and polysaccharide applied to liver organoids. The cordycepin content analyzed in the small intestine was treated with three samples of 3.14ppm, 6.28ppm, and 12.56ppm in liver organoids (differentiation 30 days, passage 2) for 24 hours, and then the size and shape of liver organoids were observed. By examining apoptosis through TUNEL assay, apoptosis of liver organoids to toxic drugs was evaluated, and cell viability was confirmed to evaluate and confirm liver organoid toxicity reactivity and predictability (Fig. 5: apoptosis result, Fig. 6). : Cell viability, y-axis represents the size of liver organoid cells and is the size ratio (%).

Summarizing Experimental Examples 1 to 3, the cordycepin-injected hydrotalcite encapsulated in Examples 1-3, that is, the cordycepin-incorporated hydrotalcite, has cordicepin inserted into the hydrotalcite bilayer structure. This worked well, and it was confirmed that the inside of the layered structure was greatly enlarged when cordycepin was inserted (Experimental Example 1). In addition, as a result of particle size measurement, the size of the hydrotalcite into which cordycepin encapsulated in Examples 1 to 3 was inserted increased by about 7 to 8 times from the average particle size of hydrotalcite of about 0.3 μm (Experimental Example 2), Comparative Example 1 Compared to 20.21 μm of , cordycepin was not easily eluted even under hydrochloric acid conditions, and when stored for a long time in the small intestine, almost 90% was eluted (Experimental Example 3). In addition, as a result of conducting a toxicity test of the eluted cordycepin, it was found that there was no toxicity in liver organoids at a maximum dose of 12.5 ppm (Experimental Example 4).

Claims (10)

removing H ₂ O and CO ₃ from hydrotalcite;
inserting cordycepin into the hydrotalcite from which the H ₂ O and CO ₃ are removed; and
Encapsulation by treating the surface of the hydrotalcite into which cordycepin is inserted with polysaccharide,
Method for preparing encapsulated cordycepin-loaded hydrotalcite.
According to claim 1,
The step of removing H ₂ O and CO ₃ from hydrotalcite is
(i) adding filtrate B and filtrate A to a basic aqueous solution, reacting, and then dehydrating to obtain a solid content;
At this time, the method for preparing the filtrate A includes adding aluminum hydroxide (Al(OH) ₃ ), NaOH, and water and heating,
The method of preparing B filtrate comprises dissolving Mg(Cl) ₂ 6H ₂ O in water.
According to claim 1,
Cordycepin is a method characterized in that cordycepin derived from Cordyceps sinensis.
According to claim 1,
In the step of inserting cordycepin into the hydrotalcite from which H ₂ O and CO ₃ are removed, the cordycepin is in the form of a material containing cordycepin, wherein the material contains 90% by weight or more of cordycepin. characterized by a method.
According to claim 1,
The method, characterized in that the polysaccharide is a polysaccharide derived from Cordyceps sinensis.
According to claim 1,
Hydrotalcite and cordycepin are used in a weight ratio of 100: 0.0001 to 0.05.
According to claim 1,
Hydrotalcite and polysaccharide are used in a weight ratio of 100:100 to 500.
The method of claim 1, characterized in that the average particle size of the encapsulated cordycepin-intercalated hydrotalcite is 0.5 to 15 μm.
A drug delivery composition comprising encapsulated cordycepin-intercalated hydrotalcite prepared by the method of claim 1.
A pharmaceutical composition comprising the composition for drug delivery of claim 9.

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